The preparation of a cover slip for cell imaging is a labor intensive operation. Typically, the preparation of the cover slip involves a number of steps for treating the cells for observation. These steps include fixating the cells to be observed for preservation and analysis. In addition, the cells are often mounted to the cover and/or immersed in a class-specific dye to enhance and highlight a specific portion of the cells for viewing. This process called “cell staining” often requires a substantial volume of dye. As is known, these dyes can be quite expensive. Once the cells have been stained according to the proper protocol, the dye must be removed or washed from the cells. Adequately washing the cells for observation may require the cells to be exposed to multiple washings. These washings can be time consuming. Given that current protocols for cell imaging contemplate a single treatment for each cover slip, it can be appreciated that even simple things like cell staining optimization require the processing of multiple cover slips.
Further, imaging multiple cover slips is a cumbersome process due to the time required to remove and replace a cover slip on a microscopic device. For example, the lens of the microscopic device must be cleaned between each use. In addition, locating the cell culture on the cover slip and properly focusing on the cells in order to provide good image contrast and high resolution may be time consuming. In view of the foregoing, it can be appreciated that it highly desirable to provide a device that reduces the time and cost associated with preparing a cover slip for cell imaging.
In order to overcome the limitations associated with preparation of a cover slip for cell imaging, various attempts have been made to develop a micro-channel system that incorporates a flow chamber that allows for the light-optical microscopic and light-spectroscopic examinations. By way of example, Kahl, United States Patent Application Publication No. US2005/0019231 A1 discloses a flow chamber made of plastic as an object carrier for light-microscopic examinations. The flow chamber includes at least one channel in a base plate. Inlet and outlet reservoirs communicate with opposite ends of the channel through an inlet and an outlet, respectively. A foil is arranged over the channel in the base plate and forms a bottom or cover for the channel. In order to make the flow chamber accessible to high-resolution microscopy, the foil is made from a relatively thin, highly transparent material. It is intended for the cells to be cultivated, fixated and stained with the channel. Thereafter, using an inverted microscope device, the cells in the channel can be observed.
While functional for its intended purpose, the flow chamber disclosed in the '231 application has certain inherent limitations. For example, the foil cannot be removed from the flow chamber and mounted on conventional cover slip. In addition, there is no mechanism for prevent leaks of fluid from the channel through the interface of the foil and the base plate. Further, the flow chamber disclosed in the '231 requires the foil to be affixed to by adhesion, hot pressing or laminating. This, in turn, increases the time associated with preparing the flow chamber for cell imaging.
Therefore, it is a primary object and feature of the present invention to provide a microfluidic device that facilitates the preparation of a sample for imaging.
It is a further object and feature of the present invention to provide a microfluidic device that facilitates the preparation of a sample for imaging that is simple and inexpensive to utilize.
It is a still further object and feature of the present invention to provide a microfluidic device that facilitates the preparation of a sample for imaging in a manner that is less time consuming than prior methods.
In accordance with the present invention, a device is proved for facilitating preparation of a sample for imaging. The device includes a cartridge having a generally flat, upper surface, a lower surface, an input port extending between the upper and lower surfaces thereof and an output port extending between the upper and lower surfaces thereof. The cartridge includes a first recess in the lower surface thereof having a first end communicating with the input port and a second end communicating with the output port. A cover slip has an upper surface engageable with the lower surface of the cartridge such that a portion of the upper surface of the cover slip communicates with the recess in the lower surface of the recess.
The device may also include a mounting structure removably connecting the cartridge to the cover slip. The mounting structure may include a substrate having an upper surface positionable against the lower surface of the cover slip and a lower surface. The substrate may includes a opening therethough for accommodating a microscopic device. The mounting structure may also include a bracket engageable with the cartridge on the substrate and a removable connector for interconnecting the bracket to the substrate and for maintaining the cartridge on the substrate. An adhesive may be provided for removably affixing the upper surface of the cover slip to the lower surface of the cartridge.
The cartridge and the cover slip may include fiduciary marks for facilitating the orientation of a microscopic device with respect thereto. In addition, the input port of the cartridge is defined by an input surface. At least a portion of the input surface is patterned to contain reagent droplets therein. It is contemplated for the outlet port in the cartridge has a generally funnel shaped cross section.
In accordance with a further aspect of the present invention, a device is provided for facilitating preparation of a sample for imaging. The device includes a cartridge having upper and lower surfaces and a first recess in the lower surface thereof. A cover slip has an upper surface engageable with the lower surface of the cartridge such that a first portion of the upper surface of the cover slip communicates with the first recess in the lower surface of the cartridge.
The lower surface of the cartridge includes a second recess therein and the upper surface of the cover slip has a second portion communicating with the second recess in the lower surface of the cartridge. The first and second recesses in the lower surface of the cartridge are generally parallel. The device may also include a substrate having an upper surface positionable against the lower surface of the cover slip and a lower surface. A removable connector is provided for maintaining the cartridge on the substrate.
The cartridge includes input and output ports. The input port is defined by a patterned input surface for containing reagent droplets therein and the outlet port has a generally funnel shaped cross section. The cartridge and the cover slip may include fiduciary marks
In accordance with a still further aspect of the present invention, a device is provided for facilitating preparation of a sample for imaging. The device includes a cartridge having upper and lower surfaces and a plurality of generally parallel recesses forming in the lower surface thereof. Each recess has an input port at a first end thereof and an output port at a second end thereof. A cover slip has an upper surface engageable with the lower surface of the cartridge such that the upper surface of the cover slip and the plurality of recesses in the lower surface of the cartridge define a plurality of channels through the device.
A substrate has an upper surface positionable against the lower surface of the cover slip and a lower surface. A removable connector maintains the cartridge on the substrate. Each input port is defined by a patterned input surface for containing reagent droplets therein. Each outlet port has a generally funnel shaped cross section. The cartridge and the cover slip include fiduciary marks.